Retracted: Study of testis histology and sperm morphology in the Bunnei,Barbus sharpeyi (Gunther, 1874) induced by luteinizing hormone‐releasing hormone analogue and carp pituitary extract

This study was conducted to determine the effects of hormone treatment on testis structure in Barbus sharpeyi, as well as the morphology of sperm examined by scanning electronic microscopy (SEM). Male B. sharpeyi were divided into three groups (three fish per group) and injected with luteinizing hormone – releasing hormone analogue (LHRH–A₂) or carp pituitary extract (CPE). The first and second groups were treated with 10 μg kg^?1 LHRH–A₂ and metoclopramide (MET), and their testis were sampled pre‐ and Poststripping respectively. The third group received 2 mg kg^?1 CPE and were killed pre‐stripping. Based on the histological results obtained, the testicular connective tissue of the lumen was thicker, and seminal vesicles were of a lower volume, in fish injected with CPE in comparison to the other groups. After treatment with LHRH–A₂ and MET, not all spermatozoa within the testis were ejaculated, and only a small amount of sperm was obtained by abdominal stripping. The highest and lowest diameters of connective tissue within lobules were observed in fish receiving CPE and LHRH–A₂ treatments respectively. There was a significant difference (P < 0.05) in lobule space between the fish injected with the CPE and the fish injected with the LHRH–A₂ and MET. The SEM results revealed that the spermatozoa of B. sharpeyi were composed of a spherical to elliptical head, a cylindrical midpiece, and a lengthy flagellum. In conclusion, it was found that injection with LHRH–A₂ and MET improved the spermatogenic process in comparison to injection with CPE.     

Fertilization rate,motility, lipid peroxidation and pH changes after chilled storage of rainbow trout (Oncorhynchus mykiss) eggs and spermatozoa by a RMPI medium

The aim of this study was to evaluate the effects of Roswell Park Memorial Institute (RPMI) 1,640 medium on chilled storage of eggs and spermatozoa of rainbow trout (Oncorhynchus mykiss). After 3 days of storage, eggs in RPMI 1,640 media (pH 8.2, 9 and 10), Cortland medium and coelomic fluid were inseminated with fresh spermatozoa (Experiment 1). Eggs in RPMI 1,640 medium at pH 8.2 shown the lowest thiobarbituric acid‐reactive substances (TBARS, 0.053 ± 0.003 nmol/ml) and pH changes (from 8.20 ± 0.01 to 8.18 ± 0.01), the highest fertilization rate (82 ± 3%). Undiluted and diluted spermatozoa at ratios of 1:2 and 1:9 with RPMI 1,640 media (pH 8.2, 9 and 10) and Cortland medium were inseminated with fresh eggs (Experiment 2). Spermatozoa in RPMI 1,640 medium at pH 9 (1:9) caused the lowest TBARS content (0.037 ± 0.002 nmol/ml) and pH changes (from 9.00 ± 0.01 to 8.98 ± 0.01), the highest fertilization rate (77 ± 2%) and motility parameters. Based on Experiments 1 and 2, eggs and spermatozoa were stored for another 3 days in RPMI 1,640 medium at pH 8.2 and 9 (1:9) respectively (Experiment 3). Fertilization rate of storage eggs and spermatozoa in Experiment 3 was 79 ± 5%, showing successfully storage of rainbow trout gametes with the same medium.     

盐度、温度和pH对2种篮子鱼精子活性的影响

选取健康成熟的雄性长鳍篮子鱼(Siganus canaliculatus)和点篮子鱼(Siganus guttatus)亲鱼进行人工采精,设定不同的盐度、温度和p H梯度,观察其对精子活力及激烈运动时间的影响。结果表明:随盐度、温度和p H的升高,2种篮子鱼精子的活力都先增强后减弱;随盐度和p H的升高精子激烈运动时间均先延长后缩短,但随温度的升高,2种篮子鱼精子激烈运动时间都呈现下降的趋势。2种篮子鱼精子的最适盐度范围均为30~35,精子活力和激烈运动都较高。长鳍篮子鱼精子最适温度范围为20~25℃,点篮子鱼精子最适温度范围为25~30℃,在25℃时,2种篮子鱼精子的活力都最高,分别为(90±2.67)%和(95±1.5)%。在p H为7~8范围内,2种篮子鱼精子活力较高,激烈运动时间也相对较长;p H为8时,长鳍篮子鱼和点篮子鱼精子活力都最高,分别为(95±4.67)%和(95±3.67)%,精子激烈运动时间也最长,分别为(44±3.89)s和(34±3.67)s。比较而言,长鳍篮子鱼精子对盐度、温度和p H的耐受性均高于点篮子鱼精子。     

Motility of sea urchin Paracentrotus lividus spermatozoa in the post‐activation phase

The characterization of sperm motility patterns, particularly post‐activation changes, is the first step in setting up species‐specific protocols involving gamete management and embryo production, for both aquaculture and laboratory research purposes. This study is aimed at the characterization of the sperm motility pattern of the purple sea urchin Paracentrotus lividus. Semen samples were individually diluted in artificial sea water for sperm motility activation. They were then incubated at 18°C for up to 24 hr. Motility was evaluated on dilution, and 1 hr, 3 hr and 24 hr after activation, by computerized analyser. The semen fertilization capacity was also evaluated. Under our experimental conditions (dilution 1:1,000 in artificial sea water plus 0.05% BSA, 18°C, in the dark), P. lividus semen remained viable for up to 24 hr, as the total motile sperm and the fertilization percentages did not change significantly during the incubation time. In contrast, the mean curvilinear velocity and the subpopulation of rapid sperm (those having a curvilinear velocity > 100 µm/s) slightly but significantly decreased after 3 hr, thereafter remaining unchanged for up to 24 hr after activation. In conclusion, our results show that diluted P. lividus semen can be used for a longer period than that of most fish species, with no need for motility inhibition procedures, supporting its wider use in laboratory research. In addition, the development of artificial fertilization protocols for aquaculture production is simplified by long‐lasting sperm motility.     

The Comet assay for detection of DNA damage in canine sperm

Sperm DNA integrity is a fundamental prerequisite in fertilization and embryo development. Among DNA integrity tests, the Comet assay is an accurate and sensitive test for the detection of sperm oxidative damage. The aim of this work was to evaluate sperm oxidative damage using the Comet assay and to study the correlation between Comet and routine assays for the evaluation of semen quality. Dogs were divided in two groups: group A (n = 6), comprising dogs with abnormal spermiogram, that is astheno‐, terato‐ or oligoasthenoteratozoospermic (OAT); and group B (n = 8), comprising normospermic dogs. The distribution of sperm oxidative damage was significantly different between the two groups (p = .001): group A—median: 31.55%, interquartile range (IQR): 30.18–38.01; group B—median: 0.90%, IQR: 0.65–1.96. The correlation between oxidative damage and abnormal morphology was high (r = .846; p < .001). There was a negative correlation between progressive motility and oxidative damage (r = ?.792; p = .001). Basal and oxidative DNA damage of spermatozoa are increased in dogs with non‐normospermic semen. In conclusion, and considering the elevated correlation with classical tests of sperm quality, the Comet assay has ample potential for clinical and research purposes in dogs.     

Effects of in vitro exposure to mercury on male gonads and sperm structure of the tropical fish tuvira Gymnotus carapo (L.)

This study investigated the progressive effects of HgCl₂ in the testis and sperm of the tropical fish tuvira Gymnotus carapo L. exposed to increasing Hg concentrations (5–30 μm ) and increasing exposure times (24–96 h). Histopathology and metal concentrations in the testis were observed. Hg concentrations in the testis reached 5.1 and 5.2 μg g^?1 in fish exposed to 20 and 30 μm of Hg, respectively. Hg effects on testicular tissue were observed even at low Hg concentrations, with no alterations in gonadosomatic index. However, the quantitative analysis of the induced alterations (lesion index) demonstrated that the Hg effects in testis became more severe after exposure to higher concentrations (20 and 30 μm ) and during longer exposure (72 and 96 h), probably leading to partial or total loss of the organ function. Hg exposure (20 μm ) also affected sperm count and altered sperm morphology. This study showed that HgCl₂ caused progressive damage to testicular tissue, reduced sperm count and altered sperm morphology. These results are important in establishing a direct correlation between Hg accumulation and severity of lesions.     

七带石斑鱼精子的超微结构

利用透射电子显微镜技术研究了七带石斑鱼（Epinephelus septemfasciatus）精子的超微结构，结果发现：七带石斑鱼精子由头部和尾部（鞭毛）两部分组成，没有明显的中片，头部前段无顶体，主要有高电子密度的球形细胞核，核中主要是致密的染色质，还分布有一些核泡，核外由核膜包围，将核与细胞质分隔开，核膜最前端紧贴质膜，核后端有植入窝，近端中心粒和中心粒间体位于其中。头部后端为袖套结构，由质膜包围的袖套结构延伸到后部，使整个头部呈椭球形，质膜后端沿鞭毛形成一个袖套腔，袖套结构中分布有中心粒复合体、线粒体以及囊泡等。精子细胞的细胞膜不光滑，呈现小的皱折。七带石斑鱼精子的尾部细长，紧接于头部细胞核，穿过袖套腔，通过基体与核紧密结合，中心粒复合体也位于此，且近端中心粒和远端中心粒呈钝角，鞭毛的轴丝由此形成，基体与鞭毛间的过渡区轴丝无中央微管，呈“9+0”型。尾部的主要结构为轴丝，其轴丝为典型的“9+2”结构，其起始端开始于前端基体，轴丝外仅有少许细胞质。     

性别控制技术在家畜育种中的应用

家畜性别控制是当今生物学领域的重大研究课题之一。近年来,性别控制技术的研究取得了可喜的进展。作者综述了国内外家畜性别控制的生物学机制、途径、方法及存在的问题和发展前景,为性别控制技术在家畜实际生产中的广泛应用提供理论基础。     

Sperm motility and seminal plasma characteristics in Barbus sharpeyi (Günther, 1874)

Spermatozoa concentration, ionic composition, osmolality, glucose and total protein contents of seminal plasma and sperm motility were determined in Barbus sharpeyi (Cyprinidae, Teleosotei). Spermatozoa concentration ranged from 9.77 to 20.20 × 10⁹ spermatozoa mL^?1. Osmolality (mOsmol kg^?1) and ionic contents (mM L^?1) of the seminal plasma were 274.5±9.0, 70.0±3.4 Na⁺, 28.8±0.9 K⁺, 101.7±3.1 Cl^?, 0.9±0.1 Mg²⁺ and 2.1±0.1 Ca²⁺ respectively. Total protein and glucose were 5.3±0.2 g L^?1 and 76.7±4.3 mM L^?1 respectively. Sperm motility was initiated in a hypo‐osmotic condition, composed of either an ionic (KCl or NaCl) or a non‐ionic (sucrose) activation medium. Duration of sperm motility was very short: <2 min after activation in distilled water. Percentage of motile spermatozoa was significantly higher in an activation medium containing NaCl compared with that of distilled water. An activating medium containing NaCl or KCl higher than 150 mM or sucrose higher than 275 mM totally inhibited the activation of sperm motility. Immediately after sperm activation, wave(s) propagated along the flagellum, but waves were restricted to the proximal part of the flagellum (close to the head) at 1 min post activation. Studied characteristics in the present study were compared with those of other cyprinids for understanding inter‐species differences.     

猪冷冻精子的体外受精与单精子胞浆内注射

用猪冷冻精子进行体外受精和单精子胞浆内注射(ICSI),并研究添加L-半胱氨酸及不同成熟培养法对猪ICSI胚胎发育的影响。结果表明:用猪冷冻精子生产的ICSI胚胎卵裂率及囊胚发育率分别为68.3%和11.4%,均低于用冷冻精子进行体外受精(IVF)所获得的卵裂率和囊胚发育率(分别为76.9%和19.2%,P<0.05)。在培养液中添加1.71mmol·L~(-1) L-半胱氨酸培养3 h,无论是卵裂率、桑椹胚发育率,还是囊胚发育率,均未得到明显改善(P>0.05);采用含0.4%BSA的NCSU-23培养液进行猪卵母细胞成熟培养,ICSI胚胎卵裂率虽明显高,但各组所获胚胎发育率均无显著差异(P>0.05),表明所用卵母细胞的成熟方法并未对ICSI胚胎发育产生明显的影响;用颗粒细胞或卵丘细胞共培养ICSI胚胎后,卵裂率有明显提高(P<0.05),但各组囊胚发育率无统计学差异。